乙型肝炎病毒共价闭合环状DNA（HBV-cccDNA）转录出多种mRNA，包括序列完整的mRNA和剪接后的mRNA，前者参与病毒功能蛋白和结构蛋白的合成；后者不参与病毒复制，但与炎症、纤维化以及肝癌的发生等有关。此外，随机整合入宿主基因的HBV-DNA片段也是mRNA的重要来源，是病毒HBsAg难以彻底清除的主要原因之一。前期工作中我们发现，在慢性乙型肝炎（CHB）自然史不同时期患者肝内HBV-mRNA水平有显著差异，但目前对出现这种差异的具体细节尚不清晰。本课题中，我们将使用三代测序技术绘制自然史中HBV全长转录组的精细图谱；利用原位核酸杂交技术直接观察肝内HBV转录本的分布以及组织学特点；通过细胞实验确定剪接RNA在调节病毒复制中的作用，以及细胞因子对HBV转录的影响。研究成果有助于进一步认识和理解HBV复制周期中的转录过程和CHB致病机理，为临床诊断和治疗提供新的科学依据。

A variety of mRNAs are transcribed from hepatitis B virus covalently closed circular (HBV cccDNA) including mRNA with intact sequence involved in the synthesis of virus functional protein and structural protein and spliced forms which are related to liver inflammation, degree of fibrosis and occurrence of liver cancer. In addition, HBV DNA fragments randomly integrate into host genome are also an important source of mRNA, which is one of the main reasons why HBsAg is difficult to be completely eliminated. In previous work, we found that there are significant differences in the levels of HBV mRNA in liver at different stages of the natural history of chronic hepatitis B (CHB).However, the details of HBV transcripts are not clear. In this proposal, we will use third-generation sequencing to fine mapping of the full length HBV transcriptome in natural history and directly visualize the distribution of viral transcripts in the liver using in situ nucleic acid hybridization. The role of spliced RNA in regulating viral replication and the effect of cytokines on viral transcription are determined by experiments of cell biology. The results of study are helpful to further understand the transcription process in the HBV life cycle and the pathogenesis of CHB, providing new proof for clinical diagnosis and treatment.